Biologia Plantarum 63: 601-609, 2019 | DOI: 10.32615/bp.2019.063

Virus-induced gene silencing for phenylalanine ammonia-lyase affects pepper adaption to low temperature

G.-X. CHENG1, J.-T. SUN1, J.-P. SHANG2, Z.-H. GONG1,*
1 College of Horticulture, Northwest A&F University, Yangling, Shaanxi 712100, P.R. China
2 Tianjin Vegetable Research Center, Tianjin 300192, P.R. China

Reverse genetics approaches in plants rely on post-transcriptional gene silencing to study the function of genes. In particular, virus-induced gene silencing (VIGS) has been successfully applied to identify gene function in some crops. To date, it is unclear whether phenylalanine ammonia-lyase (PAL) is involved in low temperature tolerance in the pepper. Here, we used an agroinfiltration protocol with tobacco rattle virus (TRV) constructs containing partial sequences from CaPAL for VIGS to test its role in anthocyanin biosynthesis and response to low temperature in the pepper (Capsicum annuum). We found that accumulation of anthocyanins in the leaves of pepper plants transformed with the TRV2:CaPAL vector was significantly reduced compared with peppers transformed with the empty TRV2 vector (TRV2:00). A significant reduction in expression of genes related to anthocyanins synthesis was also detected in peppers transformed with TRV2:CaPAL. When silenced pepper plants were exposed to a low temperature, we found decreased antioxidant system, PAL activity, and photosynthesis in plants transformed with TRV2:CaPAL compared with peppers transformed with TRV2:00. Low transcriptions of cold stress-response genes demonstrated that pepper tolerance to low temperature decreased. Future studies focused on the interaction between CaPAL and other abiotic and biotic stressors will shed further light into the role of CaPAL in stress response.

Keywords: anthocyanins, Capsicum annuum, photosynthesis, transgenic plants, TR2:CaPAL vector.

Received: June 7, 2018; Revised: March 7, 2019; Accepted: March 26, 2019; Published online: September 9, 2019Show citation

ACS AIP APA ASA Harvard Chicago IEEE ISO690 MLA NLM Turabian Vancouver
CHENG, G.-X., SUN, J.-T., SHANG, J.-P., & GONG, Z.-H. (2019). Virus-induced gene silencing for phenylalanine ammonia-lyase affects pepper adaption to low temperature. Biologia plantarum, 63, 601-609. doi: 10.32615/bp.2019.063.
Download citation
  • BibTeX (.bib)
  • Bookends (.ris)
  • EasyBib (.ris)
  • EndNote (.enw)
  • EndNote 8 (.xml)
  • ISI WoS (.isi)
  • Medlars (.medlars)
  • Mendeley (.ris)
  • MODS (.xml)
  • Papers (.ris)
  • RefWorks (.txt)
  • RefManager (.ris)
  • RIS (.ris)
  • MS Word (.xml)
  • Zotero (.ris)
    • Open full article

    Supplementary files

    Download fileCheng5844 Suppl.pdf

    File size: 383.92 kB

    References

    1. Aza-González, C., Herrera-Isidrón, L., Núñez-Palenius, H.G., Martínez De La Vega, O., Ochoa-Alejo, N.: Anthocyanin accumulation and expression analysis of biosynthesis-related genes during chili pepper fruit development. - Biol. Plant. 57: 49-55, 2013. Go to original source...
    2. Chen, J., Chen, Q., Cheng, Z.: Protective effect of anthocyanin on photosynthetic apparatus of tomato (L. esculentum Mill.) seedling leaves exposed to low temperature and low irradiance. - Sci. Agr. sin. 40: 788-793, 2007. [In Chinese]
    3. Cheng, G.X., Li, R.J., Wang, M., Huang, L.J., Khan, A., Ali, M., Gong, Z.H.: Variation in leaf color and combine effect of pigments on physiology and resistance to whitefly of pepper ( Capsicum annuum L.). - Scientia Hort. 229: 215-225, 2018. Go to original source...
    4. Cheng, G.W., Breen, P.J.: Activity of phenylalanine ammonia-lyase (PAL) and concentrations of anthocyanins and phenolics in developing strawberry fruit. - J. amer. Soc. hort. Scl. 116: 865-869, 1991. Go to original source...
    5. Christie, P.J., Alfenito, M.R., Walbot, V.: Impact of low-temperature stress on general phenylpropanoid and anthocyanin pathways: enhancement of transcript abundance and anthocyanin pigmentation in maize seedlings. - Planta 194: 541-549, 1994. Go to original source...
    6. Christopoulos, M.V., Tsantili, E.: Participation of phenylalanine ammonia-lyase (PAL) in increased phenolic compounds in fresh cold stressed walnut (Juglans regia L.) kernels. - Postharvest Biol. Technol. 104: 17-25, 2015. Go to original source...
    7. Conde, A., Chaves, M.M., Gerós, H.: Membrane transport, sensing and signaling in plant adaptation to environmental stress. - Plant Cell Physiol. 52: 1583-1602, 2011. Go to original source...
    8. Cunningham, F.X., Gantt, E.: Genes and enzymes of carotenoid biosynthesis in palnts. - Annu. Rev. Plant Physiol. Plant mol.Biol. 49: 557-583, 1998. Go to original source...
    9. Devacht, S., Lootens, P., Baert, J.V., Bockstaele, E.V., Roldan-Ruiz, I.: Influence of anthocyanin on the photosynthetic performance of industrial chicory under cold stress conditions. - Commun. Agr. appl. Biol. Sci. 74: 121-126, 2009.
    10. Dionisio-Sese, M.L., Tobita, S.: Antioxidant responses of rice seedlings to salinity stress. - Plant Sci. 135: 1-9, 1998. Go to original source...
    11. Fantini, E., Giuliano, G.: Virus-induced gene silencing as a tool to study tomato fruit biochemistry. - Methods mol. Biol. 1363: 65-78, 2016. Go to original source...
    12. Feng, H., Li, Y., Liu, Z., Liu, J.: Mapping of or, a gene conferring orange color on the inner leaf of the Chinese cabbage (Brassica rapa L. ssp. pekinensis). - Mol. Breed. 29: 235-244, 2012. Go to original source...
    13. Filiz, A., Bürün, B., Nurettin Ş.: Fungal contaminants observed during micropropagation of Lilium candidum L. and the effect of chemotherapeutic substances applied after sterilization. - Afr. J. Biotechnol. 9: 991-995, 2010. Go to original source...
    14. Guo, W.L., Chen, R.G., Gong, Z.H., Yin, Y.X., Ahmed, S.S.: Exogenous abscisic acid increases antioxidant enzymes and related gene expression in pepper ( Capsicum annuum ) leaves subjected to chilling stress. - Genet. mol. Res. 11: 4063-4080, 2012. Go to original source...
    15. Huang, J., Gu, M., Lai, Z., Fan, B., Shi, K., Zhou, Y.H., Yu, J.Q., Chen, Z.: Functional analysis of the Arabidopsis PAL gene family in plant growth, development, and response to environmental stress. - Plant Physiol. 153: 1526-1538, 2010. Go to original source...
    16. Jenshi Roobha, J., Saravanakumar, M. Aravindhan, K.M., Devi, P.S.: The effect of light, temperature, pH on stability of anthocyanin pigments in Musa acuminata bract. - Res. Plant Biol. 1: 5-12, 2011.
    17. Jeong, R.D., Hwang, S.H., Kang, S.H., Choi, H.S., Park, J.W., Kim, K.H.: Virus-induced gene silencing as tool for functional genomics in a Glycine max. - Plant Pathol. J. 21: 158-163, 2005. Go to original source...
    18. Jupin, I.: A protocol for VIGS in Arabidopsis thaliana using a one-step TYMV-derived vector. - Methods mol. Biol. 975: 197-210, 2013. Go to original source...
    19. Kim, D.S., Hwang, B.K.: An important role of the pepper phenylalanine ammonia-lyase gene (PAL1) in salicylic acid-dependent signalling of the defence response to microbial pathogens. - J. exp. Bot. 65: 2295-2306, 2014. Go to original source...
    20. Kovačević, D.B., Putnik, P., Dragović-Uzelac, V., Vahcić N., Babojelić, M.S., Levaj B.: Influences of organically and conventionally grown strawberry cultivars on anthocyanins content and color in purees and low-sugar jams. - Food Chem. 181: 94-100, 2015. Go to original source...
    21. Landi, M., Tattini, M., Gould, K.S.: Multiple functional roles of anthocyanins in plant-environment interactions. - Environ. exp. Bot. 119: 4-17, 2015. Go to original source...
    22. Leyva, A., Jarillo, J.A., Salinas, J., Martinez-Zapater, J.M.: Low temperature IiSynthase?? mRNAs of Arabidopsis thaliana in a light-dependent manner. - Plant Physiol. 108: 39-46, 1995. Go to original source...
    23. Mauch-Mani, B., Slusarenko, A.J.: Production of salicylic acid precursors is a major function of phenylalanine ammonia-lyase in the resistance of Arabidopsis to Peronospora parasitica. - Plant Cell 8: 203-212, 1996.
    24. Mittler, R., Vanderauwera, S., Gollery, M., Van Breusegem F.: Reactive oxygen gene network of plants. - Trends Plant Sci. 9: 490-498, 2004. Go to original source...
    25. Nakano, Y., Asada, K.: Purification of ascorbate peroxidase in spinach chloroplasts; its inactivation in ascorbate-depleted medium and reactivation by monodehydroascorbate radical. - Plant Cell Physiol. 28: 131-140, 1987.
    26. Olsen, K.M., Lea, U.S., Slimestad, R., Verheul, M., Lillo, C.: Differential expression of four Arabidopsis PAL genes; PAL1 and PAL2 have functional specialization in abiotic environmental-triggered flavonoid synthesis. - J. Plant Physiol. 165: 1491-1499, 2008. Go to original source...
    27. Oren-Shamir, M.: Does anthocyanin degradation play a significant role in determining pigment concentration in plants? - Plant Sci. 177: 310-316, 2009. Go to original source...
    28. Pimenta, S., Menezes, D., Neder, D.G., Melo. R.A.: Organic production systems. - Horticult. Bras. 34: 168-174, 2016. Go to original source...
    29. Pombo, M.A., Martínez, G.A., Civello, P.M.: Cloning of FaPAL6 gene from strawberry fruit and characterization of its expression and enzymatic activity in two cultivars with different anthocyanin accumulation. - Plant Sci. 181: 111-118, 2011. Go to original source...
    30. Porra, R.J., Thompson, W., Kriedemann, P.E.: Determination of accurate extinction coefficients and simultaneous-equations for assaying chlorophyll a and chlorophyll b extracted with 4 different solvents - verification of the concentration of chlorophyll standards by atomic-absorption spectroscopy. - BBA - Biomembranes 975: 384-394, 1989. Go to original source...
    31. Purkayastha, A., Mathur, S., Verma, V., Sharma, S., Dasgupta, I.: Virus-induced gene silencing in rice using a vector derived from a DNA virus. - Planta 232: 1531-1540, 2010. Go to original source...
    32. Ramegowda, V., Mysore, K.S., Senthil-Kumar, M. Virus-induced gene silencing is a versatile tool for unraveling the functional relevance of multiple abiotic-stress-responsive genes in crop plants. - Front. Plant Sci. 5: 323. 2014. Go to original source...
    33. Rohde, A.: Molecular phenotyping of the pal1 and pal2 mutants of Arabidopsis thaliana reveals far-reaching consequences on phenylpropanoid, amino acid, and carbohydrate metabolism. - Plant Cell 16: 2749-2771, 2004. Go to original source...
    34. Schulz, E., Tohge, T., Zuther, E., Fernie, A.R., Hincha, D.K.: Natural variation in flavonol and anthocyanin metabolism during cold acclimation in Arabidopsis thaliana accessions. - Plant Cell Eniron. 38:1658-1672, 2015. Go to original source...
    35. Tavakol, E. Virus-induced gene silencing (VIGS) in Aegilops tauschii and its use in functional analysis of AetDREB2. - Mol. Biotechnol. 60: 41-48, 2018. Go to original source...
    36. Tonnessen, B.W., Manosalva, P., Lang, J.M., Baraoidan, M., Bordeos, A., Mauleon, R., Oard, J., Hulbert, S., Leung, H., Leach, J.E.: Rice phenylalanine ammonia-lyase gene OsPAL4 is associated with broad spectrum disease resistance. - Plant mol. Biol. 87: 273-286, 2014.
    37. Ubi B.: Molecular mechanisms underlying anthocyanin biosynthesis: a useful tool for the metabolic engineering of the flavonoid pathway genes for novel products. - J. Food Agr. Environ. 5: 83-87, 2007.
    38. Van Tunen, A.J., Mol, J.N.M.: Control of Flavonoids Synthesis and Manipulation of Flower Color. - Springer, Dordrecht 1991. Go to original source...
    39. Wang, H., Arakawa, O., Motomura, Y.: Influence of maturity and bagging on the relationship between anthocyanin accumulation and phenylalanine ammonia-lyase (PAL) activity in 'Jonathan' apples. - Postharvest Biol. Technol. 19: 123-128, 2000. Go to original source...
    40. Zhai, Y., Wang, H., Liang, M., Lu, M.: Both silencing- and over-expression of pepper CaATG8c gene compromise plant tolerance to heat and salt stress. - Environ. exp. Bot. 141: 10-18, 2017. Go to original source...
    41. Zhao, X., Chen, T., Feng, B., Zhang, C., Peng, S., Zhang, X., Fu, G., Tao, L.: Non-photochemical quenching plays a key role in light acclimation of rice plants differing in leaf color. - Front. Plant Sci. 7: 1968, 2016.
    42. Zhou, J.X., Wang, S.Q.: Activity of phenylalanine ammonia lyase and the changes of some nitrogenous compounds in oligophenol cotton. - Plant Physiol. Commun. ??: 39-40, 1990.
    43. Zhou, L.J., Li, Y.Y., Zhang, R.F., Zhang, C.L., Xie, X.B., Zhao, C., Hao, Y.J.: The small ubiquitin-like modifier E3 ligase MdSIZ1 promotes anthocyanin accumulation by sumoylating MdMYB1 under low-temperature conditions in apple. - Plant Cell Environ. 40: 2068-2080, 2017. Go to original source...
    44. Zhu, Q., Xie, X., Lin, H., Sui, S., Shen, R., Yang, Z., Lu, K., Li, M., Liu, Y.G.: Isolation and functional characterization of a phenylalanine ammonia-lyase gene (SsPAL1) from coleus (Solenostemon scutellarioides (L.) Codd). - Molecules 20: 16833-16851, 2015. Go to original source...

    Return to the content